Effects of vibration and hyaluronic acid on activation of three-dimensional cultured chondrocytes
Effects of vibration and hyaluronic acid on activation of three-dimensional cultured chondrocytes
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DOI：
10.1002/art.21895
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发表时间：
2006-06-01
影响因子：
--
通讯作者：
Morishita, Shin
中科院分区：
文献类型：
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作者：
Takeuchi, Ryohei;Saito, Tomoyuki;Morishita, Shin
客观的。研究振动 (Vib) 和透明质酸 (HA) 对 3 维培养软骨的影响。方法。软骨细胞取自刚宰杀的 6 个月大猪的跖趾关节。使用含有 I 型胶原海绵盘的二十四孔板来培养样品。孔板振动的频率和振幅分别为100Hz和0.5nm。我们以胶原蛋白海绵为载体，利用HA和振动制作了3维软骨组织。检查了四种不同的培养条件：对照HA-Vib-组、HA-Vib+组、HA+Vib-组和HA+Vib+组。每组培养2周。培养第3、7、10和14天（每3.5天）后，测量每种培养基中合成的4-硫酸软骨素（C4S）和6-硫酸软骨素（C6S）异构体的水平。进行组织学分析、免疫组织化学分析和电镜检查。结果。平均C4S和C6S合成在培养7天后迅速增加，并且此后继续增加。 4组间差异显着（P<0.01）。 C4S和C6S的合成在HA+Vib+组中最丰富，在HA-Vib-组中最低。培养1周和2周后，各组软骨细胞均在胶原海绵上形成分层结构，但在HA+Vib+组中观察到最厚的结构，在HA-Vib-组中观察到最薄的结构。在免疫荧光下，HA+Vib+组表现出最强的色彩特征。电镜下观察HA+Vib+组软骨细胞表面出现许多细长的突起，并可见与细胞相关的细胞外物质。结论。我们的结果表明，振动和 HA 的结合激活了 3 维培养软骨细胞中蛋白聚糖的产生，并刺激 MAPK 和 β-连环蛋白。这表明软骨细胞质膜上存在一些振动机械感受器，并激活细胞内信号转导系统。
Objective. To investigate the effects of vibration (Vib) and hyaluronic acid (HA) on 3-dimensional cultured cartilage.Methods. Chondrocytes were obtained from metatarsophalangeal joints of freshly killed 6-month-old pigs. Twenty-four-well plates containing type I collagen sponge disks were used to culture samples. The frequency and the amplitude of the vibration of the well plate were 100 Hz and 0.5 nm, respectively. We produced 3-dimensional cartilage tissue using HA and vibration with collagen sponge as a carrier. Four different culture conditions were examined: a control HA-Vib- group, an HA-Vib+ group, an HA+Vib- group, and an HA+Vib+ group. Each group was cultured for 2 weeks. After culture days 3, 7, 10, and 14 (every 3.5 days), the levels of chondroitin 4-sulfate (C4S) and chondroitin 6-sulfate (C6S) isomers synthesized in each culture medium were measured. Histologic analysis, immunohistochemical analysis, and electron microscopic examination were performed.Results. Mean C4S and C6S synthesis had increased rapidly after 7 days of culture and continued to increase thereafter. There were significant differences among the 4 groups (P < 0.01). Synthesis of both C4S and C6S was most abundant in the HA+Vib+ group and the lowest in the HA-Vib- group. After I and 2 weeks of culture, the chondrocytes had formed stratified structures on the collagen sponges in all groups, although the thickest structure was observed in the HA+Vib+ group and the thinnest in the HA-Vib- group. Under immunofluorescence, the HA+Vib+ group exhibited the strongest chromatic features. Under electron microscopy, the chondrocytes in the HA+Vib+ group exhibited many long and slender prominences on their surface, and extracellular substance could be observed associated with the cells.Conclusion. Our results indicate that the combination of vibration and HA activates the production of proteoglycan in 3-dimensional cultured chondrocytes and stimulates MAPK and beta-catenin. This suggests that some mechanoreceptors for vibration exist on the plasma membrane of chondrocytes and activate the intracellular signal transduction system.